Virus Nucleic Acid Purification Reagent Kit

1. 試劑盒的組成

2. 使用說明:

1. The Virus Genome Purification Reagent Kit is intended for molecular biology research and should not be used for the prevention and treatment of diseases.
2. Prior to chemical handling, it is recommended to wear protective clothing, disposable gloves, and safety goggles.
3. Before using Rinse Buffer 1, 添加 (96%~100%) 乙醇.
4. The color change to yellow in Reagent Buffer C does not affect normal use.
5. Maintain the centrifugation environment at room temperature (15℃~25℃).

3. Reagent Kit Introduction:

The Virus Genome Purification Reagent Kit provides a rapid and effective solution for virus genome purification, widely applied to various viral nucleic acid purifications, such as Hepatitis B virus, 和更多.

The purification process can be completed within 1 小時, and the purified nucleic acids can be directly used for 聚合酶鍊式反應, 南方印跡, ETC. The entire purification process does not require toxic reagents such as phenol and chloroform, making the nucleic acid purification kit suitable for various other samples.

The purified RNA, washed with a low-salt solution or water, is suitable for downstream experiments. The A260/A280 ratio of the purified RNA is around 1.9, indicating high purity and a well-defined peak at 260 奈米.

The high-efficiency membrane effectively removes inhibitors or other interfering substances, making it widely applicable to downstream experiments such as PCR, RAPD, AFLP, RFLP, SNP, 和更多.

4. Self-provided Equipment and Consumables:

High-speed centrifuge, EP管, vortex oscillator, absolute ethanol, hot water bath or metal bath

5. Extraction Steps:

Preparation before starting:

A. Heat Elution Buffer to 65℃ before use (受到推崇的);

乙. Add ethanol to洗緩衝 1 使用前.

1. 樣品製備: Transfer 200μl of serum or other body fluids to a centrifuge tube. If insufficient, top up with PBS.
2. 添加 400µl 試劑緩衝液 C, 渦旋徹底, let it stand at room temperature for 10 分分鐘, 渦流 4-5 這段期間的次數.
3. 添加 450μl of pre-cooled absolute ethanol, vortex immediately.
4. Transfer the obtained liquid to the RNA萃取 purification column (放) (每次約650~700μl), let it stand at room temperature for 2 分分鐘, 離心機轉速超過 8,000 轉速為 1 分分鐘, 丟棄收集的廢棄物, 並將收集管重新插入純化管柱進行下一步
5. Place the RNA extraction purification column (放) 在新系列管中, 添加 300微升的 洗 緩衝 1, 離心機轉速超過 8,000 轉速為 1 分分鐘, 丟棄廢棄物, and re-insert the RNA extraction purification column (放) 進入管中進行下一步.

(筆記: Confirm that absolute ethanol has been added to 洗 緩衝 1.)

6. 添加 700µl 沖洗緩衝液 1 to the RNA extraction purification column (放), 離心機在 14,000 轉速 (20,000×g) 為了 2 分分鐘, extend the centrifugation time appropriately to ensure the membrane is more dry.

(筆記: 乙醇的存在對隨後的實驗有嚴重影響, so drying the membrane is crucial. 離心後, 確保在洗脫前不存在乙醇, then discard the waste and collection tube.)

7. Place the RNA extraction purification column (放) 放入新的離心管中, 滴 100µl 洗脫緩衝液 到膜上, 室溫孵育 5 分分鐘 (15℃~25℃), 離心機轉速超過 8,000 轉速為 1 分分鐘.

(筆記: Eluting with 50μl of elution buffer can increase nucleic acid concentration but reduces total nucleic acid yield.)

8. 重複上一步.

(筆記: A new centrifuge tube can be used to collect the nucleic acid eluted in the second round, or continue using the original collection tube to collect nucleic acid.)